Engineered microbe-targeting molecules and uses thereof
Described herein are engineered microbe-targeting or microbe-binding molecules, kits comprising the same and uses thereof. Some particular embodiments of the microbe-targeting or microbe-binding molecules comprise a carbohydrate recognition domain of mannose-binding lectin, or a fragment thereof, linked to a portion of a Fc region. In some embodiments, the microbe-targeting molecules or microbe-binding molecules can be conjugated to a substrate, e.g., a magnetic microbead, forming a microbe-targeting substrate (e.g., a microbe-targeting magnetic microbead). Such microbe-targeting molecules and/or substrates and the kits comprising the same can bind and/or capture of a microbe and/or microbial matter thereof, and can thus be used in various applications, e.g., diagnosis and/or treatment of an infection caused by microbes such as sepsis in a subject or any environmental surface. Microbe-targeting molecules and/or substrates can be regenerated after use by washing with a low pH buffer or buffer in which calcium is insoluble.
1. A composition comprising:
a. a microbe;
b. a first molecule comprising an unlabeled engineered microbe-targeting molecule which comprises a lectin or carbohydrate-binding portion thereof; and
c. a second, different, molecule comprising a detectable label conjugated to a targeting agent specific for the microbe.
2. The composition of claim 1 , wherein the engineered microbe-targeting molecule is present on a substrate.
3. The composition of claim 1 , wherein the engineered microbe-targeting molecule is covalently linked to a substrate.
4. The composition of claim 1 , wherein the lectin is a C-type lectin.
5. The composition of claim 1 , wherein the lectin is a collectin.
6. The composition of claim 1 , wherein the lectin is mannose-binding lectin.
7. The composition of claim 1 , wherein the unlabeled engineered microbe-targeting molecule comprises a carbohydrate recognition domain of mannose-binding lectin, or a fragment thereof, linked to a portion of a Fc region.
8. The composition of claim 1 , wherein the engineered microbe-targeting molecule comprises a carbohydrate recognition domain and neck region of a mannose-binding lectin fused to the C-terminal of a Fc fragment of human IgG1.
9. The composition of claim 8 , wherein the portion of the Fc region is linked N-terminal of the carbohydrate recognition domain.
10. The composition of claim 1 , wherein the lectin comprises a carbohydrate recognition domain having the sequence of SEQ ID NO: 4 or a fragment thereof.
11. The composition of claim 1 , wherein the lectin comprises a carbohydrate recognition domain having a sequence having at least 90% identity to the sequence of SEQ ID NO: 4.
12. The composition of claim 1 , wherein the detectable label comprises an enzyme.
13. The composition of claim 12 , wherein the enzyme is horseradish peroxidase (HRP).
14. The composition of claim 12 , which further comprises a substrate for the enzyme.
15. The composition of claim 14 , wherein the substrate is TMB (3,3′,5,5′-tetramethylbenzidine).
16. The composition of claim 1 , wherein the targeting agent specific for the microbe comprises an engineered microbe-targeting molecule or an antibody.
17. The composition of claim 1 , wherein the targeting agent specific for the microbe comprises MBL.
18. A kit comprising:
a. a first molecule comprising an unlabeled engineered microbe-targeting molecule which comprises a lectin or carbohydrate-binding portion thereof; and
b. one or more containers containing a second, different molecule comprising a detectable label conjugated to a targeting agent specific for a microbe.